CYTOCHEMICAL LOCALIZATION OF CELLULASE ACTIVITY IN ARTICULATED,ANASTOMOSING LATICIFERS OF PAPAVER SOMNIFERUM L. (PAPAVERACEAE)

Cellulase activity was localized at the ultrastructural level in the articulated, anastomosing laticifers of Papaver somniferum. Electron-dense crystalline deposits indicating the presence of cellulase activity were restricted to discrete patches along the laticifer wall in regions of recently formed perforations. This report presents the first direct evidence for the involvement of cellulases in the cell wall perforation process in articulated laticifers.     

Ultrastructure of spore development in <Emphasis Type="Italic">Scutellospora heterogama</Emphasis>

The ultrastructural detail of spore development in Scutellospora heterogama is described. Although the main ontogenetic events are similar to those described from light microscopy, the complexity of wall layering is greater when examined at an ultrastructural level. The basic concept of a rigid spore wall enclosing two inner, flexible walls still holds true, but there are additional zones within these three walls distinguishable using electron microscopy, including an inner layer that is involved in the formation of the germination shield. The spore wall has three layers rather than the two reported previously. An outer, thin ornamented layer and an inner, thicker layer are both derived from the hyphal wall and present at all stages of development. These layers differentiate into the outer spore layer visible at the light microscope level. A third inner layer unique to the spore develops during spore swelling and rapidly expands before contracting back to form the second wall layer visible by light microscopy. The two inner flexible walls also are more complex than light microscopy suggests. The close association with the inner flexible walls with germination shield formation consolidates the preferred use of the term ‘germinal walls’ for these structures. A thin electron-dense layer separates the two germinal walls and is the region in which the germination shield forms. The inner germinal wall develops at least two sub-layers, one of which has an appearance similar to that of the expanding layer of the outer spore wall. An electron-dense layer is formed on the inner surface of the inner germinal wall as the germination shield develops, and this forms the wall surrounding the germination shield as well as the germination tube. At maturity, the outer germinal wall develops a thin, striate layer within its substructure.     

ULTRASTRUCTURE OF SPONGIOCHLORIS TYPICA DURING SENESCENCE12

The ultrastructural changes which occurred during senescence in the stationary phase of growth of the unicellular green alga Spongiochloris typica were observed. The cell wall consists of a membrane like primary wall and an inner secondary wall which becomes progressively thickened with age of the culture. During senescence the lamellae become more compact within the chloroplast. The major feature of aging is the appearance of lipid bodies which eventually come to occupy a major portion of the cell lumen. The ultrastructural changes observed to occur during senescence are discussed in relation to physiological data.     

Morphology and ultrastructure of reproductive organs of <Emphasis Type="Italic">Monocercus arionis</Emphasis> (Sibold, 1850) Villot, 1982 (Cestoda: Cyclophyllidea)

The morphology of the reproductive system at different stages of ontogenesis and ultrastructural peculiarities of the copulatory organs and uterine in Monocercus arionis (Sibold, 1850) Villot, 1982 (Cyclophyllidea) have been studied. The muscle of the outer wall of the cirrus bag has a higher organizational level than typical smooth-muscle cells in cestodes. The cirrus is armed with typical filamentous and bladelike microtriches. The uterine epithelium contacts the thin capsule of the developing embryos located closely to the uterus wall, and the embryos contact each other in the uterine cavity in what can be interpreted as placental interactions. The specificity of the structure and arming of the copulatory apparatus has been considered, and the ultrastructural peculiarities of the uterus in the members of different orders of cestodes have been compared and analyzed.     

ULTRASTRUCTURE OF OSMOPHORES IN RESTREPIA (ORCHIDACEAE)

Osmophores of the myophilous genus Restrepia (Orchidaceae) were studied developmentally at the ultrastructural level. They are located at petal apices and on the adaxial surface of the dorsal sepal apex. Up to and through anthesis a dense, osmiophilic exudate is synthesized probably in endoplasmic reticulum or in plastids of papillose epidermal cells, and then appears to be transported through the plasmalemma by granulocrine elimination. As the exudate is amassed, the cuticle ruptures to form numerous pores that extend from the cell wall. Mitochondria and amyloplasts are especially abundant at anthesis. From anthesis to post-anthesis, lipid droplets appear in the cytoplasm and vacuoles of epidermal cells, and frequency of cell organelles drops markedly with increasing vacuolation.     

白刺小孢子和雄配子体发育的超微结构研究

对白刺小孢子和雄配子体发育的各个阶段进行了超微结构研究,结果表明在造孢细胞时期,药室内壁细胞形成“分隔细胞”;绒毛毡层细胞为分泌型,但后期完全解体而形成大量的原生质团;造孢细胞质浓,细胞器丰富;母细胞显示休眠细胞的特征;上孢子具有很厚的外壁内层;花药表皮具很厚的角质层等特征可能是旱生环境的适应。     

OBSERVATIONS ON THE RELATIONSHIP OF THE GOLGI APPARATUS TO WALL FORMATION IN THE MARINE CHRYSOPHYCEAN ALGA, PLEUROCHRYSIS SCHERFFELII PRINGSHEIM

The role of the Golgi apparatus in wall formation of vegetative cells of a marine chrysophyte, Pleurochrysis scherffelii, is described. Wall fragments are synthesized within the cisternae of the Golgi apparatus. A single Golgi apparatus is always located at the cell periphery, and the distended cisternae are oriented toward the cell surface. A highly-ordered body found near the inflated cisternae is associated with spherical, membrane-bounded bodies which may be involved in the progressive degeneration of cisternal membranes which release wall fragments. Protoplast movement has been detected by time-lapse cinephotomicrography and is correlated at the ultrastructural level with change in positions of the Golgi cisternae. Wall-synthesizing capacity is greatest during transverse wall formation. Senescent cells lack a Golgi apparatus with inflated cisternae. In addition, wall fragments are not present in the Golgi cisternae at this stage. Zoosporogenesis results in a temporary loss of the wall-forming capacity of the Golgi apparatus; this activity then resumes with the formation of a different morphological entity, the scale. Preliminary quantitative measurements of the turnover capacity of the Golgi apparatus have been made. From these data it has been determined that between 41 and 82 Golgi generations are required to synthesize the cell wall of an actively growing cell; this estimate indicates that approximately one cisterna is produced every 2 min, provided the cell generation time is 3 days. The time-lapse cinephotomicrographic data confirm that the rate of production of Golgi cisternae is at least one cisterna every 2 min.     

A procedure for the isolation of cell walls ofFucus furcatus embryos

Summary A method of isolating cell walls from Fucus embryos is described. Clean cell walls were obtained by relatively gentle means and in sufficient quantities for chemical analysis. The wall preparations appeared identical to the walls of whole embryos when compared at the ultrastructural level and by histochemical criteria.     

Xylem development and cell wall changes of soybean seedlings grown in space

BACKGROUND AND AIMS: Plants growing in altered gravity conditions encounter changes in vascular development and cell wall deposition. The aim of this study was to investigate xylem anatomy and arrangement of cellulose microfibrils in vessel walls of different organs of soybean seedlings grown in Space. METHODS: Seeds germinated and seedlings grew for 5 d in Space during the Foton-M2 mission. The environmental conditions, other than gravity, of the ground control repeated those experienced in orbit. The seedlings developed in space were compared with those of the control test on the basis of numerous anatomical and ultrastructural parameters such as number of veins, size and shape of vessel lumens, thickness of cell walls and deposition of cellulose microfibrils. KEY RESULTS: Observations made with light, fluorescence and transmission electron microscopy, together with the quantification of the structural features through digital image analysis, showed that the alterations due to microgravity do not occur at the same level in the various organs of soybean seedlings. The modifications induced by microgravity or by the indirect effect of space-flight conditions, became conspicuous only in developing vessels at the ultrastructural level. The results suggested that the orientation of microfibrils and their assembly in developing vessels are perturbed by microgravity at the beginning of wall deposition, while they are still able to orient and arrange in thicker and ordered structures at later stages of secondary wall deposition. CONCLUSIONS: The process of proper cell-wall building, although not prevented, is perturbed in Space at the early stage of development. This would explain the almost unaltered anatomy of mature structures, accompanied by a slower growth observed in seedlings grown in Space than on Earth.     

Influence of cetylpyridinium chloride on the ultrastructural appearance of sulphated glycosaminoglycans in human colonic mucosa

Summary The effect of adding cetylpyridinium chloride to the fixative on the preservation of sulphated glycosaminoglycans (SGs) was studied in human normal colonic mucosa. SGs were visualized at the ultrastructural level through the application of Spicer's High Iron Diamine (HID) technique followed by a post-fixation with potassium ferrocyanide-reduced osmium tetroxide. SGs were mainly localized in basement membranes of epithelium and capillary wall and along collagen fibers. The morphology of the reactive sites depended on the presence of cetylpyridinium chloride, SGs being granular in absence of the salt and more or less elongated when cetylpyridinium chloride was added to the fixative.We suggest that the use of cetylpyridinium chloride during fixation may help to preserve SG molecule at the ultrastructural level.     

Influence of cetylpyridinium chloride on the ultrastructural appearance of sulphated glycosaminoglycans in human colonic mucosa

The effect of adding cetylpyridinium chloride to the fixative on the preservation of sulphated glycosaminoglycans (SGs) was studied in human normal colonic mucosa. SGs were visualized at the ultrastructural level through the application of Spicer's High Iron Diamine (HID) technique followed by a post-fixation with potassium ferrocyanide-reduced osmium tetroxide. SGs were mainly localized in basement membranes of epithelium and capillary wall and along collagen fibers. The morphology of the reactive sites depended on the presence of cetylpyridinium chloride, SGs being granular in absence of the salt and more or less elongated when cetylpyridinium chloride was added to the fixative. We suggest that the use of cetylpyridinium chloride during fixation may help to preserve SG molecule at the ultrastructural level.     

Cellulase Localization in Hyphae of Achlya ambisexualis

Cellulase (EC 3.2.1.4; beta-1, 4-glucan glucanohydrolase) was localized at the ultrastructural level and found to occur in dictyosomes and vesicles, around the periphery of unidentified storage bodies, between the plasmalemma and the cell wall, and on the outer surface of the cell wall in the male strain (E87) of Achlya ambisexualis after treatment with the sex hormone antheridiol.     

Cytochemical localization of cellulase activity in pollen mother cells of david lily during meiotic prophase I and its relation to secondary formation of plasmodesmata

Summary Cellulase activity was localized at the ultrastructural level in pollen mother cells (PMCs) of David lily [Lilium davidii var.willmottiae (Wilson) Roffill] at different stages of meiotic prophase I. The enzyme was observed to appear at the early leptotene stage and reached its highest level at the subsequent zygotene stage, and its subcellular distribution revealed by the presence of electron-dense deposits of reaction product was found to be restricted exclusively to the endoplasmic reticulum (ER), the vesicles derived from that, and the cell wall, especially at the sites of secondary plasmodesmata and cytoplasmic channels where the wall was being digested. Other cytoplasmic organelles, such as dictyosomes and Golgi vesicles, lacked such deposits of reaction product. After zygotene the enzyme activity decreased abruptly, and at the pachytene stage only very few deposits could be observed in the cell wall. Our results indicate that cellulase is synthesized on rough ER and secreted directly via the smooth ER and ER-derived vesicles into the cell wall by exocytosis, where it brings about local wall breakdown, leading to the secondary formation of plasmodesmata and cytoplasmic channels.     

MORPHOLOGY,FINE STRUCTURE,AND ONTOGENY OF THE STINGING EMERGENCE OF URTICA DIOICA

Stinging emergences in Urtica dioica L. characteristically possess an elongate stinging cell and a multicellular pedestal. The emergence is derived from the epidermal and subepidermal cell layers. The apical wall of the stinging cell is composed of silica bodies which decrease basipetally in concentration. The basal portion of the cell wall of the stinging cell is devoid of silica bodies and lacks primary pit fields or pits between it and the pedestal cells. X-ray microanalysis of electron dense particles located in the stinging cell ER-golgi complex indicate that these particles contain silicon. There is no ultrastructural evidence for the presence of a toxin synthesizing system or a toxin itself.     

THE FINE STRUCTURE OF OOGENESIS IN MARCHANTIA POLYMORPHA

Archegonium development, beginning with the archegonial initial and culminating in the mature egg, was studied with the electron microscope. The ultrastructural features of the beginning stages in development of the archegonium are relatively similar to one another. Plasmodesmata occur between all adjacent cells at this time. After the secondary central cell is formed these protoplasmic connections are lost, and both axial and parietal cell lineages begin to show signs of ultrastructural differentiation. The mature egg is characterized by cytoplasm rich in ribosomes and larger organelles. Mitochondria and simplified plastids commonly display a juxtaposed association. As far as could be ascertained the numerous plastids and mitochondria in the egg of Marchantia arise through division of preexisting organelles and are not formed anew from evaginations of the nucleus. Blebbing of the nucleus produces polymorphic organelles which appear to be pinched off into the cytoplasm. The mature egg also contains vacuoles and lipid bodies toward its periphery, while dictyosomes and extensive endoplasmic reticulum occur throughout. The space between the wall cells and the mature egg appears to contain an amorphous substance. No extra membrane was observed around the mature egg.     

Polyoxin D inhibits colloidal gold-wheat germ agglutinin labelling of chitin in dimorphic forms of Candida albicans

Yeasts and mycelia of the pathogen Candida albicans grown in the presence of polyoxin D, a competitive inhibitor of chitin synthase, formed chains of swollen bulbous cells as observed by fluorescence microscopy. Wheat germ agglutinin (WGA) complexed to colloidal gold (Au) was used as a specific label at the ultrastructural level to visualize chitin in walls of control and polyoxin-treated cells. In control cells, Au-WGA labelling was preferentially localized in the innermost wall layers and was predominant at bud scars and septa. After 4.5 h in 4 mM-polyoxin D, budding in yeasts and lateral wall growth in mycelia continued, but primary septa failed to form and no Au-WGA labelling was detected in the walls. These results demonstrated that the morphological alterations caused by polyoxin D were due to the absence of chitin, a wall component important for formation of primary septa and for maintenance of structural integrity during morphogenesis.     

Quasifusulina的旋壁构造及其在(竹蜓)类分类、演化中的意义

本文介绍作者利用扫描电子显微镜对(竹蜓)类Quasifusulina旋壁超微构造的初步研究结果。电镜观察表明,Quasifusulina旋壁的主要部分系由许多近等轴的方解石微粒构成,这些微粒有规律地叠置在一起,形成了一系列垂直于壳表、彼此相邻的方解石复合体。尽管其光学特征与蜂巢层和透明层颇为相似,但从构成旋壁的方解石微粒的形状、大小及排列和定向方式来看,Quasifusulina旋壁的超微构造既明显地有别于典型的蜂巢层,也不同于典型的透明层。为便于描述,作者建议将Quasifusulina旋壁的这种特殊构造称为“原蜂巢层”(prokeriotheca)。 根据电镜观察,Quasifusulina旋壁由原蜂巢层及不连续的内疏松层构成,它可能代表了?类有孔虫中一种独特的旋壁类型。这种旋壁构造在?类动物的分类及演化研究中可能具有重要意义。     

THE THICKENED LEPTOID (SIEVE ELEMENT) WALL OF DENDROLIGOTRICHUM (BRYOPHYTA): CYTOCHEMISTRY AND FINE STRUCTURE

Leptoids (sieve elements) of Dendroligotrichum exhibit unevenly thickened lateral walls. The thickened wall areas are predominantly confined to the radial walls. With the light microscope the thickened wall cannot be resolved into distinct layers, but rather is optically homogeneous. Standard histochemical tests reveal that these walls are rich in cellulose (birefringent; IKI-H₂SO₄-positive) with small amounts of polyuronides (toluidine blue-positive) and pectins (hydroxylamine-positive) and are non-lignified. They also contain abundant natural aldehydes as revealed by the Schiff, silver hexamine, and silver proteinate reagents. Aldehyde blockades (sodium borohydrate, sodium chlorite) confirmed the presence of aldehyde groups in the cell wall. At the ultrastructural level, the lateral walls of sieve elements react strongly with uranyl and lead salts and yield little fine structural information. Electron cytochemical localization of aldehydes with silver proteinate revealed three distinct wall regions: outer, middle and inner. The outer and middle regions appeared polylamellate while the inner region contained no reaction product. The nacreous sieve elements of vascular plants are compared to the thickened sieve elements in bryophytes.